MicroRNA profiling associated with non-small cell lung cancer: next generation sequencing detection, experimental validation, and prognostic value

// Sandra Gallach 1,2 , Eloisa Jantus-Lewintre 1,2,3 , Silvia Calabuig-Farinas 1,2,4 , David Montaner 5 , Sergio Alonso 6 , Rafael Sirera 2,3 , Ana Blasco 2,7 , Marta Uso 1 , Ricardo Guijarro 8,9 , Miguel Martorell 4,10 and Carlos Camps 1,2,7,11 1 Molecular Oncology Laboratory, Fundacion Investigacion, Hospital General Universitario de Valencia, Valencia, Spain 2 Centro de Investigacion Biomedica en Red de Cancer (CIBEROnc), Madrid, Spain 3 Department of Biotechnology, Universitat Politecnica de Valencia, Valencia, Spain 4 Department of Pathology, Universitat de Valencia, Valencia, Spain 5 Department of Computational Genomics, Centro de Investigacion Principe Felipe, Valencia, Spain 6 Program of Predictive and Personalized Medicine of Cancer, Institut de Reserca Germans Trias i Pujol (PMPPC-IGTP), Badalona, Spain 7 Department of Medical Oncology, Hospital General Universitario de Valencia, Valencia, Spain 8 Department of Surgery, Universitat de Valencia, Valencia, Spain 9 Department of Thoracic Surgery, Hospital General Universitario de Valencia, Valencia, Spain 10 Department of Pathology, Hospital General Universitario de Valencia, Valencia, Spain 11 Department of Medicine, Universitat de Valencia, Valencia, Spain Correspondence to: Eloisa Jantus-Lewintre, email: // Carlos Camps, email: // Keywords : microRNAs, NSCLC, NGS, profiling, prognosis Received : May 09, 2017 Accepted : June 09, 2017 Published : June 22, 2017 Abstract Background: The average five-year survival for non-small cell lung cancer (NSCLC) patients is approximately 15%. Emerging evidence indicates that microRNAs (miRNAs) constitute a new class of gene regulators in humans that may play an important role in tumorigenesis. Hence, there is growing interest in studying their role as possible new biomarkers whose expression is aberrant in cancer. Therefore, in this study we identified dysregulated miRNAs by next generation sequencing (NGS) and analyzed their prognostic value. Methods: Sequencing by oligo ligation detection technology was used to identify dysregulated miRNAs in a training cohort comprising paired tumor/normal tissue samples ( N = 32). We validated 22 randomly selected differentially-expressed miRNAs by quantitative real time PCR in tumor and adjacent normal tissue samples ( N = 178). Kaplan-Meier survival analysis and Cox regression were used in multivariate analysis to identify independent prognostic biomarkers. Results: NGS analysis revealed that 39 miRNAs were dysregulated in NSCLC: 28 were upregulated and 11 were downregulated. Twenty-two miRNAs were validated in an independent cohort. Interestingly, the group of patients with high expression of both miRNAs (miR-21 high and miR-188 high ) showed shorter relapse-free survival (RFS) and overall survival (OS) times. Multivariate analysis confirmed that this combined signature is an independent prognostic marker for RFS and OS ( p = 0.001 and p < 0.0001, respectively). Conclusions: NGS technology can specifically identify dysregulated miRNA profiles in resectable NSCLC samples. MiR-21 or miR-188 overexpression correlated with a negative prognosis, and their combined signature may represent a new independent prognostic biomarker for RFS and OS.